Alcian Blue Staining from PBR 3383 Patient with Cystic Fibrosis

Donor Tissue Kindly Provided by Dr. Gloria Pryhuber from the University of Rochester Medical Center

1. For Frozen tissue, rinsed slides for 2X in PBS, then Incubated slides for 5 min in 4% PFA/PBS, then rinsed slides 1X in PBS.
2. Rinsed slides for 3 minutes in running dH₂O.
3. Slides were then incubated in 3% Glacial Acetic Acid (S101, Poly Scientific) for 3 minutes.
4. Slides were then incubated in filtered Alcian Blue (S111A, Poly Scientific) for 30 minutes.
5. Wash slides under cold running tap water for 10 minutes.
6. Incubate for 2 minutes in Nuclear Fast Red (S248, Poly Scientific).
7. Wash under flowing dH₂O until excess Nuclear Fast Red is washed away.
8. Incubated slides in 90% ETOH for 20 dips, repeat in different 90% ETOH.
9. Incubated slides in 100% ETOH for 20 dips, repeat in different 100% ETOH.
10. Washed slides in 2X in Xylene, followed by 6 more washes in xylene.
11. Coverslip with Permount. (Permount Toulene Solution UN 1294, Cat # SP-15-500 Fisher Scientific).

Tissue Used:
PBR 3383 Patient with Cystic Fibrosis